Effect of Bcl-x(L) Overexpression on Apoptosis and Autophagy in Recombinant Chinese Hamster Ovary Cells Under Nutrient-Deprived Condition

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Upon nutrient deprivation during Culture, recombinant Chinese hamster ovary (rCHO) cells are subjected to two types of programmed cell death (PCD), apoptosis and autophagy. To investigate the effect of Bcl-x(L) overexpression on apoptosis and autophagy in rCHO cells, an erythropoietin (EPO)-producing rCHO cell line with regulated Bcl-x(L) overexpression (EPO-off-Bcl-x(L)) was established using the Tet-off system. The expression level of Bcl-x(L) in EPO-off-Bcl-x(L) Cells was tightly regulated by doxycycline in a dose-dependent manner. Bcl-x(L) overexpression enhanced cell viability and extended Culture longevity in batch culture. Upon nutrient depletion in the later stage of batch culture, Bcl-x(L) overexpression suppressed apoptosis by inhibiting the activation of caspase-3 and -7. Simultaneously, Bcl-x(L) overexpression also delayed autophagy, characterized by LC3-II accumulation. Immunoprecipitation analysis with a Flag-tagged Bcl-x(L) revealed that Bcl-x(L), interacts with Bax and Bak, essential mediators of caspase-dependent apoptosis, as well as with Beclin-1, an essential mediator of autophagy, and may inhibit their pro-cell death function. Taken together, it was found that Bcl-x(L) overexpression inhibits both apoptosis and autophagy in rCHO cell culture. Biotechnol. Bioeng. 2009003: 757-766. (c) 2009 Wiley Periodicals, Inc.
Publisher
WILEY-BLACKWELL
Issue Date
2009-07
Language
English
Article Type
Article
Keywords

BCL-2 FAMILY PROTEINS; FED-BATCH CULTURES; CHO-CELLS; MAMMALIAN-CELLS; DOWN-REGULATION; DEATH; INHIBITION; BECLIN-1; ACTIVATION; EXPRESSION

Citation

BIOTECHNOLOGY AND BIOENGINEERING, v.103, no.4, pp.757 - 766

ISSN
0006-3592
DOI
10.1002/bit.22298
URI
http://hdl.handle.net/10203/101064
Appears in Collection
BS-Journal Papers(저널논문)
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